CN101965779B - Method for semi-lignified twig cutting breeding of Aquilaria sinensis - Google Patents

Abstract

The invention discloses a method for growing seedlings of semi-lignified twigs of agarwood. The method is to collect the seeds of Akiras chinensis from wild excellent trees, and then select excellent seedlings to build the ear-picking garden through seedling breeding; when the seedlings in the ear-picking garden grow to the second year, pruning is carried out at a distance of ≥15 cm from the ground, and the pruning is carried out. Apply 6-BA to the stems of seedlings to promote the germination of shoots; collect materials when the shoots grow semi-lignified, cut them into 10-15 cm, keep the top 3 leaves, and reserve 2/5 of the leaf area of each leaf The upper cut of the cuttings is flat, the lower end is cut obliquely and as close to the leaf nodes as possible; then the cuttings are cleaned, disinfected, and then the lower part of the cuttings is put into the prepared plant growth regulator treatment solution, and the cuttings are placed in the substrate. Manage after cutting to get Agarwood seedlings. The invention has the advantages of low cost, high survival rate, good seedling quality and fast propagation speed, can alleviate the lack of seeds, maintain the excellent genetic traits of excellent trees, is easy to popularize, and has broad development prospects.

Description

A method for growing seedlings of semi-lignified twigs of Aquilaria sinensis

technical field

The invention relates to a method for growing seedlings of semi-lignified twigs of agarwood agarwood, belonging to the technical field of plant asexual propagation.

Background technique

Aquilaria sinensis (Lour.) Gilg is a perennial tropical and subtropical evergreen tree of the genus Aquilaria in the family Daphneaceae. Agarwood is a precious medicinal plant unique to my country and an important plant resource for the production of Agarwood in my country. It is listed as a national endangered third-level protected plant and a national second-level key wild protected plant. In recent years, due to the low natural reproduction rate of Agarwood, pests and man-made deforestation, wild resources have been seriously scarce, and now they are only scattered in Hainan, Guangdong, Guangxi, Fujian and Taiwan provinces. The seed-setting rate of wild high-quality trees of Agarwood is low, it is difficult to obtain a large number of high-quality seeds, and the seeds are not suitable for preservation, and are easy to lose vitality, which makes the process of sexual reproduction management and plantation development slow and difficult.

Propagation by cuttings can solve the problems of insufficient seeds and large genetic differentiation, and open up a new way for the preservation, development and utilization of Acacia sinensis resources. Domestically, there are very few reports on the research on the cutting propagation technology of Agarwood agarwood. Only the Yunnan Academy of Forestry and the Hainan Provincial Forestry Institute have carried out preliminary research. The results show that Agarwood can be used for cuttings, but the rooting rate is very low. Systematic studies have not been reported. In recent years, the Tropical Forestry Research Institute of the Chinese Academy of Forestry, with the support of the National "Eleventh Five-Year" Science and Technology Support Project, has visited the Yunnan Academy of Forestry, the Hainan Provincial Forestry Institute and some seedling management companies to carry out cuttings of Agarwood. Seedling breeding, carried out relevant research, published a research brief (the effect of different plant growth regulators on the rooting of A. sinensis cuttings, Forestry Science Research, 2010, 23(2): 278-282), but still faced with low rooting rate and high growth rate. Seedling quality is not high. The materials used in the national patent application with the application number 200810100568.5 and the name "Method for Propagation of Baimuxiang Cuttage" and the cutting time (all year round), after repeated practice, the rooting rate of cuttings is low, and the rooting rate of cuttings in different months is low. There is a large difference in the cutting rate; the required cutting substrate is mainly moss, and the cost is high. In addition, it is required to spray the rooting accelerator within one month after the cutting, which increases the cost significantly; in terms of the survival of the cutting seedlings, only the rooting rate index is used. Due to the large limitation, the rooting quality of cutting seedlings cannot be fully measured.

Contents of the invention

The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art, and provide a kind of semi-woody agarwood with low cost, high survival rate, good seedling quality, short time required, stable and reliable, easy to popularize and can maintain the excellent properties of the mother plant. A method for raising seedlings by twig cuttings.

The object of the invention of the present invention is achieved through the following technical solutions: a method for growing seedlings of semi-lignified twigs of Aquilaria sinensis, comprising the following steps:

(1) Preparation of panicle seedlings: collect the Akiras chinensis seeds of wild excellent trees, sow and raise seedlings, then select excellent seedlings, and build ear picking gardens; in the year of the bamboo season, the seedlings in the ear picking gardens are carried out at a distance of not less than 15cm from the ground. Pruning, smearing 6-BA (6-benzylaminopurine) on the pruned seedling stems to promote germination of shoots; when the shoots grow semi-lignified, materials can be collected;

(2) Preparation of the substrate: fully mix the yellow heart soil, peat soil and vermiculite in a volume ratio of 3:2: (1-2) to obtain the substrate;

(3) Preparation of cuttings: in the morning (7:00-9:00) and when the light is low (≤10000lux), take the semi-lignified shoots obtained in step (1) and cut them into 10-15cm , keep the top 3 leaves, and keep 2/5 of the leaf area of each leaf; the upper cut of the cuttings is flat to reduce water loss, and the lower end is cut obliquely and as close to the leaf nodes as possible; then the cuttings are cleaned and disinfected, and then the lower part of the cuttings is cut Put it into the prepared plant growth regulator treatment solution, and then cut it into the substrate;

(4) Post-cutting management: after the cutting is completed, the temperature of the cutting environment is maintained at 15-30° C. and the relative humidity is not less than 75%; at the same time, the cutting environment is sterilized; and the cutting seedlings of Agarwood sinensis are obtained.

The method for raising seedlings by cuttings of semi-lignified twigs of Aquilaria edulis more preferably includes the following steps: after obtaining the cutting seedlings of Agarwood edulis, the rooting rate, the number of hairy roots, the longest root length and partial The root rate is comprehensively evaluated, and the cutting seedlings of Agarwood with good quality are obtained; the calculation equation for the membership function value [X(u1), X(u2)] used for the comprehensive evaluation is:

X(u 1)＝[XX _min ]/[X _max -X _min ] (1)

X(u2)＝1-[XX _min ]/[X _max -X _min ] (2)

Wherein, X is the measured value of a certain evaluation index; X _max is the maximum value of all data obtained by a certain evaluation index; X _min is the minimum value of all data obtained by a certain evaluation index; the evaluation index includes rooting rate , the number of hair roots, the longest root length and partial root rate; finally, the membership functions of all evaluation indicators are added up;

If the measured evaluation index is positively correlated with the quality of cutting seedlings, formula (1) is used to calculate the membership value, otherwise, formula (2) is used; the specific membership values of each evaluation index of each treatment are added up, and the greater the cumulative value, the obtained Agarwood. The better the quality of the cuttings.

The wild excellent tree described in step (1) is preferably a wild tree whose diameter at breast height is not less than 60cm and tree height is not less than 20m;

The excellent seedlings described in step (1) are preferably seedlings of 10 to 12 months with a diameter of not less than 0.6 cm and a seedling height of not less than 60 cm;

The concentration of 6-BA (6-benzylaminopurine) described in step (1) is preferably 100 mg·L ⁻¹ ;

The substrate described in the step (2) is preferably placed in a long 540mm, wide 280mm, deep 110mm specification 50 hole seedling dish, and the potassium permanganate solution of 0.3% by mass percentage is used for 2 days before the cutting to carry out sprinkling and disinfection of the substrate, The substrate is permeable; the 50-hole seedling tray with a length of 540mm, a width of 280mm, and a depth of 110mm has the advantage of being easy to install the substrate, saving space, and the roots are clustered together after the seedlings grow, which is convenient for afforestation and improves the survival rate;

The month of preparation of cuttings described in step (3) is preferably April;

The cleaning method described in step (3) is preferably rinsing in running water for 5 minutes;

The disinfection described in the step (3) is preferably to be sterilized with 0.1% potassium permanganate solution by mass percentage for 1min, and then rinsed with distilled water;

The plant growth regulator treatment solution described in step (3) is preferably IAA (indole acetic acid) or IBA (indole butyric acid); more preferably the IAA whose concentration is 2500mg L ^-1 or the concentration is 1500mg L ^{- 1} IBA;

The time for putting the lower part of the cuttings into the prepared plant growth regulator treatment solution as described in step (3) is preferably quick dipping for 8 to 15 seconds;

The cuttage method described in step (3) is preferably a straight insertion method, that is, the cuttings are vertically inserted into the plug matrix, and one cutting is cut in each hole;

The method of cutting described in step (3) is more preferably to use bamboo strips to make a guide hole on the matrix before cutting, the depth is 3-4cm, and then insert the cuttings vertically into the matrix of the plug tray, and cut 1 cutting in each hole ;After inserting, slightly compact the substrate around it and water thoroughly;

The specific method of disinfection described in the step (4) is to carry out alternate disinfection with 0.1% potassium permanganate solution and 1000 times of carbendazim every 3 to 4 days; after 20 days, alternate disinfection at this concentration , once a week.

The present invention has following advantage and effect:

The invention has the advantages of low cost, high survival rate, good seedling quality and fast propagation speed, and can solve the problem of seed shortage. In addition, the selected materials are wild seedlings of excellent trees, and the method can maintain the excellent properties of the mother plant. Generally speaking, the present invention is easy to popularize and has broad development prospects.

Detailed ways

The present invention will be further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.

Example 1

(1) Preparation of panicle seedlings: collect Agarwood seeds (source is Hainan Province) of wild excellent trees (diameter at breast height ≥ 60cm, tree height ≥ 20m), carry out seed seedling cultivation, and then select excellent seedlings (diameter ≥ 0.6cm, height ≥ 20m) ≥60cm) to build an ear-picking garden; when the seedlings grow to the second year, prune at a distance ≥15cm from the ground, and apply 6-BA to the pruned seedling stems to promote germination of shoots; when the shoots grow semi-lignified can be collected when available.

(2) Selection and processing of cuttings: in April, choose the morning (7:00～9:00) and when the light is low (≤10000lux), take the 2-year-old seedlings germinated in the ear picking nursery obtained in step (1). For semi-lignified branches, cut them into 10-15cm long, keep the top 2-3 leaves, and keep 2/5 of the leaf area of each leaf; the cuttings are cut flat to reduce water loss, and the lower end is cut obliquely and as close as possible At the leaf node.

(3) Substrate treatment and container selection: fully mix yellow heart soil, peat soil, and vermiculite in a volume ratio of 3:2:1 to obtain a substrate; Potassium manganate solution is sprayed for disinfection, and the matrix shall prevail. The cuttage container used in this operation is a 50-hole seedling tray with a length of 540mm, a width of 280mm, and a depth of 110mm. Its advantages are that it is easy to install the substrate and save space.

(4) Cutting and post-cutting management: Dissolve IAA or IBA in 20 mL of absolute ethanol, and then make 200 mL of IAA (2500 mg·L ^-1 ) or IBA (1500 mg·L ^-1 ) treatment solution. Rinse the cuttings prepared in step (2) in running water for 5 minutes, take them out and put them in a 0.1% potassium permanganate solution for disinfection for 1 minute, rinse them with distilled water, and then put the lower part of the cuttings into the previously prepared IAA Or in the solution for IBA treatment, quickly dip for 10s, and cut the cuttings after the alcohol evaporates. The straight insertion method is adopted, that is, the cuttings are inserted vertically into the matrix of the hole plate, and one cutting is cut in each hole. In order to prevent the cuttings from being damaged during cutting, use bamboo strips to make a guide hole in the substrate before cutting, with a depth of 3-4cm. After cutting, slightly compact the substrate around it and water it thoroughly. This operation is cutting in the greenhouse. After the cutting is completed, a small shed is built on the seedbed and covered with a plastic film to keep the humidity of the cutting environment. Most of the strong light in the greenhouse has been filtered out by the greenhouse shading net, but it is still necessary to use a shading net with a light transmittance of 35% for shading at noon when the sun is sufficient to prevent excessive transpiration of the cuttings leaves, and the cuttings lose water and die. dry. After cuttings, alternate disinfection with 0.1% potassium permanganate solution and 1000 times carbendazim every 3 to 4 days; after 20 days, alternate disinfection at this concentration, once a week; at the same time, spray every day , Control the air humidity above 80%, with the upper part of the cuttings moist and fresh. Three replicates were performed.

(5) At the same time, set up a control experiment (three repeated experiments each):

A group contrast is to adopt the semi-lignified twig branch that germinated in the same year of non-2-year-old seedlings, and other steps are with embodiment 1;

Group B contrast is different seedling cultivation containers, and other steps are with embodiment 1;

C group contrast is different cuttage positions, and other steps are with embodiment 1;

D group contrast is the matrix of different formulations, and other steps are with embodiment 1;

Group E is contrasted with different cutting seasons, and other steps are the same as in Example 1;

Among them, the control materials of group B to group E were all 2-year-old semi-lignified shoots that germinated in the same year, and the specific experimental design is shown in Table 1:

Table 1

(6) Results: after cutting by the method of the present invention, on the 7th day after inserting, the lenticels at the incision of the cuttings were enlarged and accompanied by white flocculent callus; after 14 days, the lenticels of the incisions of most of the cuttings were enlarged and cracked At the same time, more callus appeared, connecting the phloem and xylem of the incision; on the 20th day of cutting, the callus of the cuttings increased to the most, and some even covered the entire incision; on the 35th day after cutting, the cuttings were 0.1cm away from the upper part of the incision There are white bud-like adventitious roots in the area of ~0.5cm; on the 50th day after cutting, the cuttings take root one after another. In each control group, after 10 days of cutting, calluses were produced successively in the incision of the cuttings, and adventitious roots were produced after 40 days.

When cutting for 6 months, the rooting quality indicators (rooting rate, hairy root number, longest root length, partial root rate) of cutting plants were investigated, and the rooting rate, hairy root number, longest root length and The partial root rate was used for comprehensive evaluation of cutting seedlings. The comprehensive evaluation value of cutting seedlings adopting the technical measures of the present invention is that the membership function value is up to 3.04, and its rooting rate can reach more than 75%, the number of hairy roots is greater than 10 ears ^-1 , the longest root length is more than 12.3cm, and partial root The rate is less than 25%, and the quality of seedlings is significantly higher than that of each control group. The results of each control group are shown in Table 2.

Table 2

Note: The membership function method, the calculation equation of the membership function value [X(u1), X(u2)] used for analysis is:

X(u1)＝[XX _min ]/[X _max -X _min ] (1)

X(u2)＝1-[XX _min ]/[X _max -X _min ] (2)

Wherein, X is the measured value of a certain evaluation index; Xmax is the maximum value of all data obtained by a certain evaluation index; Xmin is the minimum value of all data obtained by a certain evaluation index; root number, longest root length and partial root rate; finally, add up the membership functions of all evaluation indicators; if the measured indicators are positively correlated with the quality of cutting seedlings, use (1) to calculate the membership value, otherwise use (2) formula; accumulating the specific subordination values of each index for each treatment, the greater the accumulative value, the better the quality of the obtained Agarwood seedlings.

The rooting quality of the cuttings is comprehensively evaluated by the membership function method, and the method adopted in the present invention has a high membership function value, and the cutting seedlings are transplanted, and the survival rate is as high as 95% or more, and the later growth is robust.

The present invention uses common yellow heart soil, peat soil and vermiculite as the cutting matrix, and has low cost, and the cutting container can be reused, which saves a lot of cost. The cuttage of Aquilaria sinensis through this method has good rooting quality, can be rooted 35 days after cutting, can be transplanted in 6 months, requires a short time, can maintain good properties, is stable and reliable, and is easy to popularize.

The above-mentioned embodiment is a preferred embodiment of the present invention, but the embodiment of the present invention is not limited by the above-mentioned embodiment, and any other changes, modifications, substitutions, combinations, Simplifications should be equivalent replacement methods, and all are included in the protection scope of the present invention.

Claims (9)

1. the method for a buta-buta semi-lignified epicormic branch cutting is characterized in that comprising following steps:

(1) preparation of fringe seedling: gather the buta-buta seed of wild select tree, carry out seed seedling-raising, select good seedling construction cutting orchard again; When growth of seedling to the second year, prune being not less than the 15cm place apart from ground, smear 6-benzyl aminopurine at the seedling stem place of pruning with the short spray of sprouting; Treat to draw materials when the spray growth is semi-lignified;

(2) preparation of matrix: with yellow soil, peat soil, vermiculite 3: 2 by volume: (1～2) fully mixes, and obtains matrix;

(3) insert the preparation of fringe: select 7:00～9:00 in morning, when illumination is not more than 10000lux, get the semi-lignified spray that step (1) obtains, it is cut grow into 10～15cm, keep 3 blades in top, every leaf keeps 2/5 leaf area; Inserting the fringe upper cut and be flat mouthful, cuts sth. askew and locates near leaf segment as far as possible in the lower end; Then clean and insert fringe, carry out disinfection, will insert the fringe bottom again and put into the plant growth regulator treatment fluid for preparing, and then cuttage is in matrix;

(4) manage after the cuttage: after cuttage finished, 15～30 ℃ of cuttage environment maintenances, relative moisture were not less than 75%, and the cuttage environment is carried out disinfection, and obtain the buta-buta cuttage seeding;

Plant growth regulator treatment fluid described in the step (3) is heteroauxin or indolebutyric acid;

The concentration of said heteroauxin is 2500mgL ^-1

The concentration of said indolebutyric acid is 1500mgL ^-1

2. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting; It is characterized in that: said method also comprises following steps: the long and inclined to one side root rate of rooting rate, root of hair quantity, the longest root that obtains through Subordinate Function buta-buta being inserted fringe again after the buta-buta cuttage seeding is carried out overall merit, obtains superior in quality buta-buta cuttage seeding; Membership function value [X (u1), X (the u2)] accounting equation that is used for overall merit is:

X(u1)＝[X-X _min]/[X _max-X _min] (1)

X(u2)＝1-[X-X _min]/[X _max-X _min] (2)

Wherein, X is the measured value of certain evaluation index; X _MaxThe maximum of all data that obtain for certain evaluation index; X _MinThe minimum of a value of all data that obtain for certain evaluation index; Described evaluation index comprises rooting rate, root of hair quantity, the longest root is long and inclined to one side root rate; Membership function with all evaluation indexes adds up at last;

If institute the valency index of test and assess and cuttage seeding quality are proportionate, employing (1) formula calculating membership values then, on the contrary then use (2) formula; Add up and respectively handle the concrete membership values of each evaluation index, accumulated value is big more, and the quality of the buta-buta cuttage seeding that obtains is good more.

3. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting, it is characterized in that: wild select tree described in the step (1) refers to the diameter of a cross-section of a tree trunk 1.3 meters above the ground and is not less than the wild-type tree that 60cm, the height of tree are not less than 20m.

4. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting, it is characterized in that: good seedling described in the step (1) refers to leading thread and is not less than 10～12 months the seedling that 0.6cm, height of seedling are not less than 60cm.

5. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting, it is characterized in that: the concentration of 6-benzyl aminopurine is 100mgL described in the step (1) ^-1

6. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting; It is characterized in that: the matrix described in the step (2) is placed long 540mm, wide 280mm, dark 110mm specification 50 cave seedling dishes; Preceding 2 days liquor potassic permanganates with mass percent 0.3% of cuttage drench matrix irritates sterilization; Be as the criterion with the matrix of drenching, and then be used for cuttage.

7. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting, it is characterized in that:

The preparation month of the said slotting fringe of step (3) is April;

Cleaning the mode of inserting fringe described in the step (3) is that fringe 5min is inserted in flushing in flowing water;

Sterilization described in the step (3) is to insert fringe 1min with the disinfecting solution of potassium permanganate of mass percent 0.1%, and is clean with distilled water flushing again;

To insert the fringe bottom described in the step (3), to put into the time of the plant growth regulator treatment fluid for preparing be that speed is dipped in 8～15s;

The mode of cuttage is the straight cutting method described in the step (3).

8. according to the method for claim 7 buta-buta semi-lignified epicormic branch cutting, it is characterized in that:

The mode of cuttage described in the step (3) is on matrix, to make a call to a guiding hole with the bamboo bar earlier before the cuttage, and the degree of depth is 3～4cm, will insert fringe again and vertically insert in the disk base material of cave, and fringe is inserted in cuttage 1 strain in every cave; After inserting with the compacting and water permeable a little of matrix around it.

9. according to the method for claim 1 buta-buta semi-lignified epicormic branch cutting, it is characterized in that:

The concrete grammar of sterilization described in the step (4) is that liquor potassic permanganate and 1000 times of carbendazim of 0.1% replace sterilization for every mass percent of using respectively at a distance from 3～4 days; After 20 days, by the alternately sterilization of this concentration, 1 time weekly.